Gut microbiome influenced many aspects of host physiology and psychology. Vice versa, lifestyles factors such as exercise and healthy diet are ways to shape the gut microbiota towards balance. We observed two distinct microbe groups characterized by physical fitness in a multi-omic cohort of 2183 young subjects with metagenomics, national physique comprehensive test, lifestyle and metabolome data. The panel of bacterial taxa including Clostridium bolteae, Escherichia coli, Ruminococcus gnavus, Clostridium clostridioforme, Clostridium innocuum, Bacteroides cellulosilyticus and Oscillospiraceae, were consistently associated with most of the physical fitness. Clostridium species and trace element both increased in the individuals those tend to stay up late. Yogurt consumption was associated with Streptococcus thermophilus and Bifidobacterium animalis subsp. lactis in feces, which differed from potentially endogenous Bifidobacterium species that was associated with milk intake. Our large-scale analyses were poised to advise for a healthy gut microbiome through behavioural changes.